5V2X

Ethylene forming enzyme in complex with manganese and 2-oxoglutarate

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	100
Detector technology	CCD
Collection date	2016-10-16
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	0.9787
Spacegroup name	P 21 21 21
Unit cell lengths	74.411, 97.546, 102.385
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	44.032 - 1.847
R-factor	0.1645
Rwork	0.163
R-free	0.20080
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5v2t
RMSD bond length	0.007
RMSD bond angle	0.750
Data reduction software	XDS
Data scaling software	Aimless (0.5.28)
Phasing software	PHASER (2.7.16)
Refinement software	PHENIX (1.11.1-2575)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	45.330	45.330	1.890
High resolution limit [Å]	1.847	9.050	1.847
Rmerge	0.131	0.030	0.744
Rmeas	0.145	0.033	0.820
Rpim	0.060	0.015	0.341
Number of reflections	64351
<I/σ(I)>	11.6
Completeness [%]	99.6	98.5	95.9
Redundancy	5.8	4.8	5.6
CC(1/2)	0.997	0.999	0.830

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8	277.15	0.18 ul 61 mg/ml EFE (25 mM HEPES pH 8.0, 1 mM TCEP, 0.6 mM 2-oxoglutarate) was mixed with 0.18 ul reservoir solution. The sitting drop reservoir of 50 ul contained 0.2 M lithium chloride, 0.1 M Tris-HCl pH 8.0, 20% w/v Polyethylene glycol 6,000. The crystal was soaked for about a minute in 25% glycerol, 75% reservoir solution before freezing it.